Reddy S, Bibby N J, Smith P A, Elliott R B
Aust J Exp Biol Med Sci. 1985 Dec;63 ( Pt 6):667-81. doi: 10.1038/icb.1985.70.
A convenient procedure is described for the purification of rat trypsin. Tissue was homogenized and extracted at pH 4 and the soluble fraction purified by a two-step affinity chromatography. After polyacrylamide gel electrophoresis at pH 8.8, the purified enzyme was resolved into 1 major and 2 minor bands all of which possessed trypsin-like enzyme activity. Antibodies to rat trypsin were raised in rabbits and utilized in establishing a sensitive radioimmunoassay procedure for the enzyme. The assay was adapted to study the levels of the enzyme in the circulation of normal Wistar and spontaneously diabetic Bio Breeding Wistar rats before onset of insulin-dependent diabetes mellitus in a longitudinal fashion. In the normal rat, serum levels of immunoreactive trypsin were higher in younger animals and showed a decline after weaning. This pattern was also seen in the Bio Breeding Wistar rats. In about half the number of Bio Breeding Wistar rats, serum immunoreactive trypsin levels were much higher than in normal rats. These results may imply that in some Bio Breeding Wistar rats the disease may be associated with inflammatory lesions of the exocrine pancreas.
本文描述了一种简便的大鼠胰蛋白酶纯化方法。将组织在pH 4条件下匀浆并提取，可溶性部分通过两步亲和层析进行纯化。在pH 8.8条件下进行聚丙烯酰胺凝胶电泳后，纯化的酶被分离为1条主要条带和2条次要条带，所有条带均具有胰蛋白酶样酶活性。用兔制备大鼠胰蛋白酶抗体，并用于建立该酶的灵敏放射免疫分析方法。该分析方法经调整后用于纵向研究正常Wistar大鼠和自发糖尿病Bio Breeding Wistar大鼠在胰岛素依赖型糖尿病发病前循环中该酶的水平。在正常大鼠中，免疫反应性胰蛋白酶的血清水平在年幼动物中较高，断奶后呈下降趋势。这种模式在Bio Breeding Wistar大鼠中也可见。在大约一半数量的Bio Breeding Wistar大鼠中，血清免疫反应性胰蛋白酶水平远高于正常大鼠。这些结果可能意味着在一些Bio Breeding Wistar大鼠中，该疾病可能与外分泌胰腺的炎症性病变有关。
